Terrestrial Ecotoxicity of Eight Chemicals in a Systematic Approach (7 pp)

Background and Objective   Terrestrial ecotoxicity data are required for many research purposes. The data are derived either from the literature or elaborated by own investigations. As the terrestrial toxicity tests are usually time-consuming and labour intensive, the experiments are performed with a limited number of test organisms and soils. In the context of a project sponsored by CEFIC-LRI (European Chemical Industry Council – Long-Range Research Initiative), EC50-values were systematically elaborated for eight chemicals with a wide range of logKow-values (CdCl2, Trinitrotoluene, 3,4-dichloroaniline, 2,4-dichlorophenol, Tributyltinchloride, Pentachlorophenol, Benzo(a)pyrene, p,p-dichloro-2,2-diphenyl-1,1,1-trichloroethane). The substances were selected covering a broad range of physico-chemical and ecotoxicological properties. As toxicity endpoints, microbial activities, plant germination and growth as well as reproduction of earthworms and collembola were determined. As such systematic investigations are rarely performed and for some substances no data existed, the data pool is made available to the scientific community.Methods   All toxicity tests were conducted on three different soil types (sandy soil, silty soil, loamy soil), according to ISO and OECD guidelines Results, Discussion, Conclusion and Outlook   The different toxicities of the chemicals, the influence of soil properties on bioavailability as well as different sensitivities of test organisms and test parameters are reflected by the EC50-values. The results showed that the EC50-values calculated on the basis of nominal concentrations can significantly vary from EC50-values derived from analytical concentrations for some substances. To avoid false conclusions, this has to be considered especially when concentrations determined in the field are compared with toxicity data obtained from the literature or calculated on the basis of nominal concentrations. Moreover, the results indicate that terrestrial ecotoxicological tests should be accompanied by chemical analyses, to be sure to derive sound EC50-values. As the tests were performed according to international guidelines, the comprehensive data pool relevant for the terrestrial ecotoxicity of eight chemicals is considered to be of great value for further research and discussions dealing with the habitat function of soils.     

Presence and dispersal of infective Frankia in peat and meadow soils in Sweden

Summary Use of the N₂-fixing grey alder, Alnus incana (L.) Moench, as a short-rotation crop for energy production is currently being explored. To evaluate the need for inoculation of alders, the distribution of infective propagules of Frankia in the soil at potential sites for alder plantations was examined. Uninoculated grey alder seedlings were grown in three types of soil. Frequent nodulation was found in a meadow soil which had been free from actinorhizal plants for nearly 60 years, but the alder seedlings failed to nodulate in peat soil from two different bog sites. One of these bogs had been exploited for peat and the surface layer of the peat had been removed, so that the soil samples were taken from deep layers of the peat. At the other site, an area of cultivated peat, there were no infective propagules of Frankia in plots without alders; the infective Frankia was present in plots only where it had been introduced by inoculated alders. There was no detectable air-borne dispersal of Frankia. Instead, water movement might account for the dispersal of Frankia in peat. Although the apparent absence of Frankia in these peat soils necessitates inoculation of alder seedlings before planting out, this makes it possible to introduce and maintain Frankia strains with selected beneficial characteristics, since there is no competition from an indigenous Frankia flora.     

Estimates of denitrification in soil by remote sensing of thermal infrared emission at different moisture levels

Remote sensing techniques may be one way to narrow the range of uncertainty in extrapolating N₂ emissions from small-scale to large-scale terrestrial ecosystems. In the present work we investigated the correlations between denitrification activity, soil moisture, and soil thermal infrared emissions. A field experiment was performed on two different agricultural soils, one loam and one silty clay. The results indicated that thermal infrared emissions can only be used to estimate the denitrification rate in soil within a limited range of soil moisture levels. Estimates of denitrification activity based on soil texture and moisture are, however, very likely to be a fruitful approach to generating large-scale N fluxes.     

Effects of tetracycline on the soil microflora: function,diversity, resistance

Background  Tetracycline is a widely used antibiotic in animal production. Significant amounts of the substance reach the soil via feces, urine and manure application. As tetracycline is a persistent compound with antibacterial activity, its presence in soil may have undesired direct and indirect effects. These have been investigated so far focusing on effects on selected microbial functions. Objectives  The aim of the present study was to obtain comprehensive information on potential effects of tetracycline on the soil microflora under environmentally relevant conditions. The investigations included function and structure of the microbial biocoenosis and the distribution of resistance genes. Methods  Pig manure rich in tetracycline resistance genes was applied to a sandy soil. This soil as well as an unamended soil were additionally treated with several concentrations of tetracycline. The spiked soils were incubated in outdoor lysimeters for several months. Substrate induced respiration, PLFAs, ten selected resistance genes, and the concentrations of tetracycline were determined. Results  The test concentrations, though far exceeding environmental relevance, caused only small effects. An establishment of resistance could not be detected. Applied resistance genes were not detectable at the end of the study even in the presence of added tetracycline. Conclusion  Due to the high sorption capacity of the antibiotic, environmentally relevant concentrations of tetracycline do not seem to cause undesired effects on the soil microflora.     

Are forests potential landscape barriers for foraging bumblebees? Landscape scale experiments with Bombus terrestris agg. and Bombus pascuorum (Hymenoptera, Apidae)

Habitat fragmentation is generally considered to have detrimental effects on insect movement and associated pollen flow between plant populations. Against this background the role of forests as potential barriers for foraging bumblebees of the species Bombus terrestris agg. and Bombus pascuorum was studied in different experiments. Bombus terrestris agg. workers were found foraging at distances of up to 2.2 km from their nests. B. terrestris agg. as well as B. pascuorum individuals crossed 600 m of forestland between floral mass resources (Phacelia tanacetifolia, Helianthus annuus), although in general a high degree of site fidelity was observed. B. pascuorum workers accepted artificial floral resources within the forest, whereas the minor use of resources below forest canopy observed for B. terrestris agg. possibly indicates a preference for direct forage flights, probably leading above the forest canopy. Our results warn against experiments with genetically modified crop species with potential bumblebee pollination (e.g. Brassica napus), in which an isolating effect of forests is assumed.     

The Application of Multivariate Statistical Methods for the Evaluation of Soil Profiles (8 pp)

Background, Aim and Scope   Contamination of soils does not only occur on their surface over large areas, but also in depth. Therefore a characterization of soil state after pollution demands a three-dimensional soil sampling, by what a large number of samples has to be analyzed. Analytical results could be evaluated by multivariate statistical methods, which have already been used for the evaluation of data sets containing results from soil sampling of two dimensions like areas or single profiles. In this case study, multivariate statistical methods were applied to investigate structure and interactions between features in a data set containing results of three-dimensional soil sampling. The investigated soil profiles were contaminated by emissions of a former cement and phosphate fertilizer plant. The aim of this study was to determine the remaining extent of contamination and to analyze whether pollutants are mobilized and vertically transported within the profiles. Materials and Methods: Three soil profiles were sampled in the surroundings of the plant. Grain size, organic and carbonatic bonded carbon, pH value, and the total contents of Ca, Cd, Co, Cu, F, Fe, K, Mn, Mg, Na, Ni, P, Pb, and Zn were determined. The resulting data set was evaluated by cluster analysis, linear discriminant analysis, and principal components analysis. The sequential extraction procedure according to Zeien and Brümmer was applied to analyze the binding properties of Ca, Cd, Cu, Na, Pb, and Zn from selected samples. Results: Cd was identified as contaminant of the top soils. The pH values of the bottom soils were determined to be in alkaline range, which is unnaturally high. Variables were clustered according to enrichment of variables in top soils. The samples were classified regarding their pollution state and their substrate by cluster analysis, which was confirmed by linear discriminant analysis. Geogenic and anthropogenic sources of variables as well as relationships between variables like the binding of heavy metals at organic matter were detected by using principal components analysis. The binding of heavy metals at organic matter in the top soils was confirmed by the results of the applied sequential extraction. A vertically altered distribution of Na binding was determined. Discussion: According to the current soil conditions, the uptake of heavy metals had probably occurred by the over ground part of plants during the deposition. The distribution of Na should likely result from the vertical transport of Na, which would also explain the high pH values of the bottom soils by ion exchange. Altogether, the main amount of deposited Ca, F, Na, P, and heavy metals is likely nearly insoluble bound in the top soils. Conclusions: Ten years after the end of production, the pollution of top soils in the surroundings of the former plant is still high. However, regarding the ecotoxicological relevance the now explored interactions between several soil features and elements strongly indicate that there is no short-term to medium-term risk of a mobilization of the deposited elements with the exception of Na. Recommendations and Perspectives: The results of this case study prove that multivariate statistical methods are powerful tools to explore interactions of variables and relationships in a data set derived from three dimensional soil sampling. The methods applied in this work can be highly recommended for evaluations of large data sets resulting from two- or three-dimensional samplings. Multivariate statistical methods enable the characterization of soils and their pollution state in a simple and economic way.     

Pathogenicity determinants in smut fungi revealed by genome comparison

Biotrophic pathogens, such as the related maize pathogenic fungi Ustilago maydis and Sporisorium reilianum, establish an intimate relationship with their hosts by secreting protein effectors. Because secreted effectors interacting with plant proteins should rapidly evolve, we identified variable genomic regions by sequencing the genome of S. reilianum and comparing it with the U. maydis genome. We detected 43 regions of low sequence conservation in otherwise well-conserved syntenic genomes. These regions primarily encode secreted effectors and include previously identified virulence clusters. By deletion analysis in U. maydis, we demonstrate a role in virulence for four previously unknown diversity regions. This highlights the power of comparative genomics of closely related species for identification of virulence determinants.     

Effects of pulsation rate and viscosity on pulsation-induced taste enhancement: new insights into texture-taste interactions

Oral stimulation with high-tastant concentrations that are alternared with low-tastant concentrations or water rinses (pulsatile stimulation) results in taste intensity ratings that are higher than continuous stimulation with the same average tastant concentration. This study tested the combined effects of taste pulsation rate and viscosity on pulsation-induced taste enhancement in apple juice. According to a tastant-kinetics hypothesis, less pulsation-induced taste enhancement is expected at enhanced pulsation rates in the high-viscous proximal stimulus compared to lower viscous stimuli. High-concentration sucrose apple juice pulses and low-concentration sucrose apple juice intervals were alternated at different pulsation periods (pulse + interval in seconds) every 2.5 s (period length = 5 s) or every 1.25 s (period length = 2.5 s). Pulsed stimuli were presented at two viscosity levels by the addition of pectin (0 and 10 g/L). Sweetness intensities of pulsed stimuli were compared to a continuous reference of the same net but nonalternating sucrose concentration. Sweetness ratings were higher for pulsatile stimuli than for continuous stimuli. In low-viscous stimuli, enhancement depended on the pulsation period and peaked at 5 s periods. In high-viscous stimuli, the same enhancement was observed for both pulsation periods. These results contradict a tastant-kinetics hypothesis of viscosity-induced taste suppression because impaired tastant kinetics by viscosity would predict the opposite: lower pulsation-induced taste enhancement for viscous stimuli, especially at higher pulsation rates. Instead, these observations favor an explanation based on perceptual texture-taste interactions, which predict the observed independence between viscosity and pulsation rate.     

Time lag between peak concentrations of plasma and salivary cortisol following a stressful procedure in dairy cattle

Background

Measurement of salivary cortisol has been used extensively as a non-invasive alternative to blood sampling to assess adrenal activity in ruminants. However, there is evidence suggesting a considerable delay in the transfer of cortisol from plasma into saliva. Previous studies in cattle have used long sampling intervals making it difficult to characterise the relationship between plasma and salivary cortisol (PLCort and SACort, respectively) concentrations at different time points and determine whether or not such a time lag exist in large ruminants. Therefore, the objective of this study was to characterise the relationship between plasma and salivary cortisol and determine if there is a significant time lag between reaching peak cortisol concentrations in plasma and saliva across a 4.25 h time-period, using short sampling intervals of 10–15 min, following social separation in dairy cattle.Five cows were separated from their calves at 4 days after calving, and six calves were separated from a group of four peers at 8 weeks of age. Following separation, the animals were moved to an unfamiliar surrounding where they could not see their calves or pen mates. The animals were catheterised with indwelling jugular catheters 1 day before sampling. Blood and saliva samples were obtained simultaneously before and after separation.

Results

In response to the stressors, PLCort and SACort increased reaching peak concentrations 10 and 20 min after separation, respectively. This suggested a 10 min time lag between peak cortisol concentrations in plasma and saliva, which was further confirmed with a time-series analysis. Considering the 10 min time lag, SACort was strongly correlated with PLCort (P < 0.0001).

Conclusions

Salivary cortisol correlates well with plasma cortisol and is a good indicator of the time-dependent variations in cortisol concentrations in plasma following acute stress. However, there is a time lag to reach peak cortisol concentrations in saliva compared to those in plasma, which should be considered when saliva samples are used as the only measure of hypothalamic-pituitary-adrenal axis response to stress in cattle.     

MAGNETIC RESONANCE IMAGING OF PULP IN NORMAL AND DISEASED EQUINE CHEEK TEETH

A retrospective analysis of 619 upper and lower cheek teeth from 62 horses was performed. Based on clinical findings, as well as radiographic and magnetic resonance (MR) imaging findings, the teeth were classified into five groups. There were 20 teeth with abnormal MR imaging signals as well as clinical alterations and 599 healthy teeth. Using MR imaging, the appearance of pulp in diseased and disease‐free teeth was compared, and the appearance of abnormal pulp was studied. Subsequently, the ability of MR imaging to diagnose pulpitis and pulp necrosis in teeth with normal external appearance was investigated. In horses with clinically verified dental disease, abnormal MR imaging findings were confirmed in the pulp of all affected teeth. An enlarged blurred pulp image with a lower signal intensity was observed only in clinically diseased teeth and was a reliable criterion for diagnosing dental disease on MR imaging. On the other hand, partial or complete absence of pulp in all MR imaging sequences was observed in both diseased and nondiseased teeth. These data demonstrate that pulp changes in equine cheek teeth can be evaluated using MR imaging.